Experimental Techniques Research Articles

PSMD9 promotes the malignant progression of hepatocellular carcinoma by interacting with c-Cbl to activate EGFR signaling and recycling

BackgroundMounting evidences shows that the ubiquitin‒proteasome pathway plays a pivotal role in tumor progression. The expression of 26S proteasome non-ATPase regulatory subunit 9 (PSMD9) is correlated with recurrence and radiotherapy resistance in several tumor types. However, the role and mechanism of PSMD9 in hepatocellular carcinoma (HCC) progression remain largely unclear.MethodsPSMD9 was identified as a prognosis-related biomarker for HCC based on analysis of clinical characteristics and RNA-seq data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) and the JP Project of the International Cancer Genome Consortium (ICGC-LIRI-JP). PSMD9 expression was analyzed in cancer tissues and adjacent noncancerous tissues via immunohistochemistry and Western blotting. Multiple in vivo and in vitro experimental techniques (such as CCK-8, colony formation, EdU, and Transwell assays; flow cytometry; Western blotting; quantitative RT-PCR; Coimmunoprecipitation assay and immunofluorescence confocal imaging) were used to assess the functions of PSMD9 in the pathogenesis of HCC.ResultsWe found that the expression of PSMD9 was upregulated and associated with a poor prognosis in HCC patients. PSMD9 promoted HCC cell proliferation, migration, invasion and metastasis. Knockdown of PSMD9 significantly inhibited HCC cell proliferation by inducing G1/S cell cycle arrest and apoptosis. Mechanistically, we demonstrated that PSMD9 promoted HCC cell proliferation and metastasis via direct interaction with the E3 ubiquitin ligase c-Cbl, suppresses EGFR ubiquitination, influenced EGFR endosomal trafficking and degradation and subsequently activated ERK1/2 and Akt signaling. In addition, we showed that PSMD9 knockdown sensitized HCC cells to the tyrosine kinase inhibitor erlotinib in vitro and in vivo.ConclusionsCollectively, our results indicate that PSMD9 drives HCC progression and erlotinib resistance by suppressing c-Cbl mediated EGFR ubiquitination and therefore can be a potential therapeutic target for HCC.

Dynamics of Protein-RNA Interfaces Using All-Atom Molecular Dynamics Simulations.

Facing the current challenges posed by human health diseases requires the understanding of cell machinery at a molecular level. The interplay between proteins and RNA is key for any physiological phenomenon, as well protein-RNA interactions. To understand these interactions, many experimental techniques have been developed, spanning a very wide range of spatial and temporal resolutions. In particular, the knowledge of tridimensional structures of protein-RNA complexes provides structural, mechanical, and dynamical pieces of information essential to understand their functions. To get insights into the dynamics of protein-RNA complexes, we carried out all-atom molecular dynamics simulations in explicit solvent on nine different protein-RNA complexes with different functions and interface size by taking into account the bound and unbound forms. First, we characterized structural changes upon binding and, for the RNA part, the change in the puckering. Second, we extensively analyzed the interfaces, their dynamics and structural properties, and the structural waters involved in the binding, as well as the contacts mediated by them. Based on our analysis, the interfaces rearranged during the simulation time showing alternative and stable residue-residue contacts with respect to the experimental structure.

Electronic Effect of the [Au(PPh3)]+ Fragment in the Stabilization of Imidazolium Salts and the Destabilization of NHCs

AbstractThis work presents the synthesis and characterization of mono‐ (1) and di‐nuclear (2) imidazolium salts stabilized by [AuPPh3]+ fragments. The presence of the gold moiety induces a significant decrease in the carbenic proton‘s acidic character (high pKa). This high stability is consistent with the pronounced instability of the conjugate bases obtained through deprotonation using strong bases. The formation of carbene species is accompanied by the identification of a 1,2‐rearrangement process in which a preference for the C‐bound species over the N‐bound species is observed. Experimental techniques such as NMR, single‐crystal X‐ray diffraction analysis, mass spectrometry, and computational calculations are employed to investigate the reactivity exhibited by the imidazolium salts. Additionally, the electrochemical properties of the two imidazolium salts were also investigated. This study reveals that both species 1 and 2 display two cathodic peaks which are related to two electro‐chemical irreversible reduction events. The results obtained from both experimental and theoretical studies of this system reveal a strong tendency of the [AuPPh3]+ fragment to stabilize imidazolium salts. Additionally, they demonstrate the preference of this fragment for C‐bound species over N‐bound ones, with the former proving to be highly unstable even under severe conditions of air and moisture exclusion.

Advancements in P3HT:PCBM solar cells through experimental and simulated techniques

Abstract Organic solar cells are a promising alternative in photovoltaics due to their low manufacturing cost and flexibility. In this study, we developed an organic solar cell of structure (ITO/PEDOT:PSS/P3HT:PCBM/Al) where the active layer is represented by P3HT:PCBM. Experimental studies revealed that this layer has an energy gap of 1.9 eV and a good absorption coefficient of 〖3×10〗^5 〖cm〗^(-1). Using analytical and experimental methods based on (J-V-T) curves, we extracted the key parameters of our device, including parasitic resistances (R_s and R_sh), ideality factor (n), and Schottky potential barrier (ϕ_B) at different temperatures. Also the charge transport mechanism in our junction is identified in this study. We then used simulation to evaluate the performance of our solar cell, obtaining an initial efficiency of 2.13%, in line with experimental results from other research. Our results confirm the feasibility of our theoretical approach, with performance consistent with previous work. However, by introducing an ETL layer (PFN-Br or PDINO), we were able to improve efficiency up to 4.25%. These results highlight the importance of optimising intermediate layers to maximise the efficiency of organic solar cells.

Image‐based inertial impact test for viscoelastic constitutive identification: A digital replica for error quantification

AbstractPolymers find widespread use in applications where they are subjected to impact loading. Therefore, understanding the time dependence of their mechanical response is critical to the design of structures subjected to these high strain rate environments. However, characterising these materials on microsecond time scales has proven challenging. Traditional experimental techniques rely on satisfying a number of limiting assumptions and typically do not provide direct measurements of the material parameters. Here, we propose a novel implementation of the image‐based inertial impact (IBII) test to extract viscoelastic constitutive parameters on these microsecond time scales using the stress gauge implementation of the virtual fields method. We validate the experiment using a digital replica approach in which the constitutive parameters are first extracted on a finite element model of an IBII test on a viscoelastic material. The finite element data are then used to synthetically deform computer‐generated grid images, which are then polluted with grey‐level noise to simulate the images that would be captured in a real‐life experiment. These images are processed identically to a physical experiment, and the identification is repeated using the full‐field displacements extracted from the computer‐generated images to determine the ideal processing parameters. Parameter identification was found to strongly depend on the processing parameters used to extract the kinematic fields from full‐field images, emphasising the need for computational validation before attempting a physical experiment to extract constitutive parameters. The IBII experimental method was found to be capable of simultaneously identifying the bulk modulus and the shear modulus along with their associated time constant.

Cyclic R‐Curve Measurements for Structural Metallic Alloys

Fractures nucleated from defects and subjected to cyclic loading can experience propagation for a range of stress intensity factor ΔK well below the so‐called long crack threshold. This phenomenon is attributed to the development of crack closure mechanisms which may differ from those observed in laboratory tests conducted in accordance with current standards. Cracks originating from material defects require a specific degree of extension to develop the plastic wake, thus achieving a stabilized condition called the long crack threshold. However, in certain materials, this stabilization length can extend up to several millimeters, effectively encompassing a significant portion of the component's fatigue life. Therefore, understanding and quantifying the development of ΔKth with crack extension is important for implementing a reliable assessment procedure based on the fracture mechanics theory. Herein, R‐curve measurements are presented for five distinct structural metallic alloys widely used in various industrial applications. Additionally, the impact of load ratio is investigated, providing a comprehensive analysis of fatigue crack resistance in terms of the R‐curve concept. This study underscores the necessity of ruling new experimental techniques to measure and implement the long crack threshold, thereby ensuring the development of a reliable and robust framework for fatigue assessment.

Regulation and Role of Adiponectin Secretion in Rat Ovarian Granulosa Cells

Adiponectin is an important adipokine involved in glucose and lipid metabolism, but its secretion and potential role in regulating glucose utilization during ovarian development remains unclear. This study aims to investigate the mechanism and effects of follicle-stimulating hormones (FSHs) on adiponectin secretion and its following impact on glucose transport in the granulosa cells of rat ovaries. A range of experimental techniques were utilized to test our research, including immunoblotting, immunohistochemistry, immunofluorescence, ELISA, histological staining, real-time quantitative PCR, and transcriptome analysis. The immunohistochemistry results indicated that adiponectin was primarily located in the granulosa cells of rat ovaries. In primary granulosa cells cultured in vitro, both Western blot and immunofluorescence assays demonstrated that FSH significantly induced adiponectin secretion within 2 h of incubation, primarily via the PKA signaling pathway rather than the PI3K/AKT pathway. Concurrently, the addition of the AdipoR1/AdipoR2 dual agonist AdipoRon to the culture medium significantly stimulated the protein expression of GLUT1 in rat granulosa cells, resulting in enhanced glucose absorption. Consistent with these in vitro findings, rats injected with eCG (which shares structural and functional similarities with FSH) exhibited significantly increased adiponectin levels in both the ovaries and blood. Moreover, there was a notable elevation in mRNA and protein levels of AdipoRs and GLUTs following eCG administration. Transcriptomic analysis further revealed a positive correlation between the expression of the intraovarian adiponectin system and glucose transporter. The present study represents a novel investigation, demonstrating that FSH stimulates adiponectin secretion in ovarian granulosa cells through the PKA signaling pathway. This mechanism potentially influences glucose transport (GLUT1) and utilization within the ovaries.

Transient Temperature at Tool–Chip Interface during Initial Period of Chip Formation in Orthogonal Cutting of Inconel 718

Machining nickel-based super alloys such as Inconel 718 generates a high thermal load induced via friction and plastic deformation, causing these alloys to be among most difficult-to-cut materials. Localized heat generation occurring in machining induces high temperature gradients. Experimental techniques for determining cutting tool temperature are challenging due to the small dimensions of the heat source and the chips produced, making it difficult to observe the tool–chip interface. Therefore, theoretical analysis of cutting temperatures is crucial for understanding heat generation and temperature distribution during cutting operations. Periodic heating and cooling occurring during cutting and interruption, respectively, are modeled using a hybrid analytical and finite element (FE) transient thermal model. In addition to identifying a transition distance associated with initial period of chip formation (IPCF) from apparent coefficient of friction results using a sigmoid function, the transition temperature is also identified using the thermal model. The model is validated experimentally by measuring the tool–chip interface temperature using a two-color pyrometer at a specific cutting distance. Due to the cyclic behavior in interrupted cutting, where a steady-state condition may or may not be achieved, transient thermal modeling is required in this case. Input parameters required to identify the heat flux for the transient thermal model are obtained experimentally and the definitions of heat-flux-reducing factors along the cutting path are associated with interruptions and the repeating IPCF. The thermal model consists of two main parts: one is related to identifying the heat flux, and the other part involves the determination of the temperature field within the tool using a partial differential equation (PDE) solved numerically via a 2D finite element method.

What Fechner could not do: Separating perceptual encoding and decoding with difference scaling.

A key question in perception research is how stimulus variations translate into perceptual magnitudes, that is, the perceptual encoding process. As experimenters, we cannot probe perceptual magnitudes directly, but infer the encoding process from responses obtained in a psychophysical experiment. The most prominent experimental technique to measure perceptual appearance is matching, where observers adjust a probe stimulus to match a target in its appearance along the dimension of interest. The resulting data quantify the perceived magnitude of the target in physical units of the probe, and are thus an indirect expression of the underlying encoding process. In this paper, we show analytically and in simulation that data from matching tasks do not sufficiently constrain perceptual encoding functions, because there exist an infinite number of pairs of encoding functions that generate the same matching data. We use simulation to demonstrate that maximum likelihood conjoint measurement (Ho, Landy, & Maloney, 2008; Knoblauch & Maloney, 2012) does an excellent job of recovering the shape of ground truth encoding functions from data that were generated with these very functions. Finally, we measure perceptual scales and matching data for White's effect (White, 1979) and show that the matching data can be predicted from the estimated encoding functions, down to individual differences.

Experimental and computational study on antioxidant activity and molecular properties of novel ferrocenyl Schiff bases

In this study, antioxidant and molecular properties of some important novel ferrocenyl Schiff bases were studied. The 4-Ferrocenyl aniline synthetic pathway was chosen as the general procedure for the synthesis of Schiff bases. Two experimental techniques, UV–Visible spectroscopy and cyclic voltammetry, and one theoretical DFT method was employed to find out their antioxidant activity and molecular properties of some novel ferrocenyl Schiff bases and compared it with standard antioxidant (Ascorbic acid). Maximum wavelength, extinction coefficient, IC30 and IC50 were estimated by using UV–Visible spectroscopy. For theoretical investigations, computational Amsterdam density function software was employed to study the molecular properties and reaction parameters based on density functional theory calculations. The cyclic voltammetry (CV) measurements in electrochemical experiments were done at room temperature while diffusion coefficient was estimated by using CV graphs. UV–Vis spectra of these compounds exhibited similar peaks with greater intensity ranges from 312 nm to 314 nm and smaller intensity ranges 471 nm to 473 nm which occurred because of the only substituent R (naphthoxy, phenoxy and phenoxy) in these compounds. Registered Amsterdam density function was utilized to determine the optimized structure, FTIR spectra, UV–Visible spectra, HOMO, LUMO, dipole moment, ionization potential, electron affinity, band gap, hardness, chemical potential and electrophility index. The overall electrode processes were found to be diffusion controlled and the novel compounds showed appreciable antioxidant activities.

LAMP2A regulates cisplatin resistance in colorectal cancer through mediating autophagy

BackgroundDrug resistance is an important constraint on clinical outcomes in advanced cancers. LAMP2A is a limiting protein in molecular chaperone-mediated autophagy. This study was aimed to explore LAMP2A function in cisplatin (cis-diamminedichloroplatinum, DDP) resistance colorectal cancer (CRC) to seek new ideas for CRC clinical treatment.MethodsIn this study, LAMP2A expression was analyzed by molecular experimental techniques,such as qRT-PCR and western blot. Then, LAMP2A in cells was interfered by cell transfection experiments. Subsequently, the function of LAMP2A on proliferation, migration, invasion, DDP sensitivity, and autophagy of CRC/DDP cells were further investigated by a series of experiments, such as CCK-8, transwell, and western blot.ResultsWe revealed that LAMP2A was clearly augmented in DDP-resistant CRC and was related to poor patient prognosis. Functionally, LAMP2A insertion remarkably CRC/DDP proliferation, migration, invasion ability and DDP resistance by strengthen autophagy. In contrast, LAMP2A knockdown limited the proliferation, migration, and invasion while heightened cellular sensitivity to DDP by restraining autophagy in CRC/DDP cells. Furthermore, LAMP2A silencing was able to curb tumor formation and enhance sensitivity to DDP in vivo.ConclusionIn summary, LAMP2A boosted malignant progression and DDP resistance in CRC/DDP cells through mediating autophagy. Clarifying LAMP2A function in DDP resistance is promising to seek cancer therapies biomarkers targeting LAMP2A activity.

Microbial Fuel Cells Performance Evaluation through the Application of Fuzzy Logic

Objective: This work aimed to employ fuzzy logic combined with the design of experiments technique to statistically evaluate how the microbial fuel cells operating parameters influence their performance. Theoretical Framework: Microbial fuel cells (MFC) are a technology of interest in the current scenario as they allow simultaneously promoting the biotreatment of waste and the bio generation of electrical energy. Methodology: Through a bibliographical search based on publications on Google Scholar platform over the last 10 years, it was noticed that anode area, external electrical resistance and reactor volume are the most reported input parameters in MFC research and current density and power density are the output parameters most frequently portrayed in these studies, which is why these variables were selected for statistical investigation. Results and Discussion: The results showed that, for both outputs studied, reactor volume and anode area showed a positive effect, while the external electrical resistance showed a negative effect. It was also possible to develop mathematical models that indicated the relationship between the input and output variables studied, with statistical significance for power density model (R2 = 86%). Originality: Applying computational simulations and subsequent design of experiments to obtain results in accordance with those obtained experimentally in the laboratory.

Non-equivalent Atomic Vibrations at Interfaces in a Polar Superlattice.

In heterostructures made from polar materials, e.g., AlN-GaN-AlN, the non-equivalence of the two interfaces has long been recognized as a critical aspect of their electronic properties, in that they host different two-dimensional carrier gases. Interfaces play an important role in the vibrational properties of materials, where interface states enhance thermal conductivity and can generate unique infrared-optical activity. The non-equivalence of the corresponding interface atomic vibrations, however, has not been investigated so far due to a lack of experimental techniques with both high spatial and high spectral resolution. Herein we experimentally demonstrate the non-equivalence of AlN-(Al0.65Ga0.35)N and (Al0.65Ga0.35)N-AlN interface vibrations using monochromated electron energy-loss spectroscopy in the scanning transmission electron microscope (STEM-EELS) and employ density-functional-theory (DFT) calculations to gain insights in the physical origins of observations. We demonstrate that STEM-EELS possesses sensitivity to the displacement vector of the vibrational modes as well as the frequency, which is as critical to understanding vibrations as polarization in optical spectroscopies. The combination enables direct mapping of the non-equivalent interface phonons between materials with different phonon polarizations. The results demonstrate the capacity to carefully assess the vibrational properties of complex heterostructures where interface states dominate the functional properties. This article is protected by copyright. All rights reserved.

Extra-articular hip resection with maintenance of pelvic continuity in malignant tumours of the proximal femur with articular invasion.

Extra-articular hip resection may be necessary in cases of malignant tumour of the pelvic bone or of the proximal femur invading the hip joint. When the tumour is in the proximal femur, it is possible to resect the acetabulum en bloc by performing a periacetabular osteotomy, but this creates a discontinuity in the pelvic ring with difficult reconstruction and diminished function. Several techniques described recently seek to be as sparing as possible on the pelvic bone by preserving the posterior column or both columns in order to facilitate reconstruction and improve function. However, these still require complex reconstructions and can necessitate intra-pelvic dissection. We describe here an extra-articular hip resection technique for tumours of the proximal femur invading the joint, with maintenance of pelvic continuity by preserving both columns and the quadrilateral plate of the acetabulum, without intra-pelvic dissection, that can be performed on patients in whom the medial wall of the acetabulum is thick enough. Our preliminary assessments have included studies on dry bone and imaging analyses. The technique was first tested on a single cadaver pelvis (encompassing 2 hips) and subsequently performed on a patient with a pathological fracture of the femoral neck due to osteosarcoma secondary to Paget's disease. Further clinical applications are essential to evaluate the overall effectiveness, safety and impact on patient functionality of this experimental technique.

USP31 Activates the Wnt/β-catenin Signaling Pathway and Promotes Gastric Cancer Cell Proliferation, Invasion and Migration.

Gastric cancer (GC) has a poor prognosis because it is highly aggressive, yet there are currently few effective therapies available. Although protein ubiquitination has been shown to play a complex role in the development of gastric cancer, to date, no efficient ubiquitinating enzymes have been identified as treatment targets for GC. The TCGA database was used for bioinformatic investigation of ubiquitin-specific protease 31 (USP31) expression in GC, and experimental techniques, including Western blotting, qRT-PCR, and immunohistochemistry, were used to confirm the findings. We also analyzed the relationship between USP31 expression and clinical prognosis in patients with GC. We further investigated the effects of USP31 on the proliferation, invasion, migration, and glycolysis of GC cells in vitro and in vivo by using colony formation, CCK-8 assays, Transwell chamber assays, cell scratch assays, and cell-derived xenograft. Furthermore, we examined the molecular processes by which USP31 influences the biological development of GC. Patients with high USP31 expression have a poor prognosis because USP31 is abundantly expressed in GC. Therefore, USP31 reduces the level of ubiquitination of the Wnt/β-catenin pathway by binding to β-catenin, thereby activating glycolysis, which ultimately promotes GC proliferation and aggressive metastasis. USP31 inhibits ubiquitination of β-catenin by binding to it, stimulates the Wnt/β-- catenin pathway, activates glycolysis, and accelerates the biology of GCs, which are all demonstrated in this work.

Mechanical Properties of Mouse Lung Cells and Their Effects on the Atomic Force Microscope Beam Vibrations.

In the present investigation, the mechanical properties of mouse normal and carcinomatous (LL/2) lung tissue cells were investigated using atomic force microscopy (AFM). The normal lung cells have been derived directly from C57BL mice. Initially, the elastic modulus of LL/2 cells was measured following chemotherapy with the anti-cancer drug Cisplatin and plasma treatment. MTT evaluation was used to determine the optimal dosages for 24- and 48-h incubations based on the IC50 cell viability concentration during chemotherapy treatment. After 24 and 48 h, the results demonstrated that Cisplatin-based chemotherapy increases the elastic modulus of LL/2 cells by 1.599 and 2.308 times compared to untreated cells. LL/2 cells were subsequently treated with plasma for 30 and 60 s for 24 and 48-h incubation. The plasma treatment decreased the LL/2 cell's elastic modulus, and the time duration of plasma treatment increased the reduction amount of elastic modulus. During the second section of the study, theoretical (finite element analysis [FEM]) and experimental techniques were used to examine the resonant frequencies and magnitude of the frequency response function (FRF) of the AFM cantilever's movements when applying normal and cancerous cells before and after chemo and plasma treatments as specimens. The results indicated that increasing the samples' elastic modulus raises the resonant frequency, so the resonant frequency of treated cells as a sample is greater than untreated cells. In conclusion, the FEM and experimental results were compared and found to be in good agreement.

Thermal decomposition of oxygen‐containing Ta3N5${\rm {Ta}}_3{\rm {N}}_5$

AbstractTransition metal nitrides, especially tantalum nitrides, are pivotal for applications in extreme environments demanding excellent mechanical properties and thermodynamic stability. Among them, ‐TaN, a high‐pressure polymorph of tantalum nitride with its exceptional bulk modulus (362 GPa) and hardness (31.7 GPa) promises to have many technological uses. Another nitride, , has gained importance as a photocatalyst for water splitting using visible light. The Ta–N phase diagram indicates that the thermal decomposition of pure leads to the formation of ‐TaN. However, usually has some amount of oxygen as an impurity mainly due to its synthesis route. We found that the ‐TaN phase, which is usually observed at high pressures, is formed during the thermal decomposition of oxygen containing . The presence of ‐TaN is verified using several experimental techniques such as X‐ray diffraction, Raman spectra, high‐angle annular dark field scanning transmission electron microscopy (STEM‐HAADF), and electron energy loss spectroscopy (EELS). Elemental distribution analyzed through energy dispersion X‐ray spectroscopy (XEDS) in STEM reveals about 7 at.% of oxygen in ‐TaN. First‐principle calculations are performed to examine the thermodynamic stability of oxygen substituted ‐TaN and pure ‐TaN via formation enthalpies, elastic constants, and phonon dispersion calculations. The computational studies confirm that oxygen in ‐TaN enhances its thermodynamic stability. The calculated electron localization functions establish the bonding characteristics between Ta, N, and O, confirming the same.

High-fidelity single logical qubit encoding scheme assisted by single-sided quantum dot-cavity systems.

We present an encoding scheme of a single logical qubit with single-sided quantum dot (QD)-cavity systems, which is immune to the collective decoherence. By adjusting the Purcell factor to satisfy the balanced reflection condition, the detrimental effects of unbalanced reflection between the coupled and uncoupled QD-cavity systems can be effectively suppressed. Furthermore, the fidelity of each step can be increased to unity regardless of the strong coupling regime and the weak coupling regime of cavity quantum electrodynamics (QED) with the assistance of waveform correctors. The scheme requires QD-cavity systems and simple linear optical elements, which can be implemented with the currently experimental techniques.

Improved implementation of concentrated plasticity models in real-time hybrid simulation

Real-time hybrid simulation (RTHS) is a cost-effective experimental testing technique for evaluating the dynamic response of structural systems. In RTHS, a portion of the structure is modeled numerically (i.e., numerical substructure), while the remaining part is tested physically (i.e., experimental substructure). These substructures are coupled in real time. A significant challenge in RTHS is accurately computing the dynamic response of the numerical substructure in real time during testing. Constrained by this challenge, numerical substructures in RTHS are often oversimplified (e.g., spring-mass-dashpot models), failing to capture more complex dynamic behaviors and limiting the applicability of RTHS. However, with the increasing use of innovative structural systems, new materials, and advanced design guidelines to ensure resilient and sustainable civil infrastructure against natural hazards, the demand for real-time executable higher-fidelity numerical substructures in RTHS is growing. Consequently, concentrated plasticity model (CPM) has gained attention as the numerical substructure in RTHS for its good balance between numerical accuracy and computational efficiency, as well as its popularity in structural engineering applications. Currently, static condensation is utilized to prevent an increase in computational degrees of freedom (DOFs) induced by the hinges in CPM, aiming to save computational cost. However, this method proves counterproductive for nonlinear analyses. While reducing the number of computational DOFs, it complicates the stiffness representation, thereby necessitating more demanding arithmetic operations for stiffness updating. In this study, multi-point constraints are applied to enhance the computational efficiency, specifically for nonlinear analyses involving stiffness updating, and the versatility of CPM. Moreover, direct stiffness updating method and sparse matrix-vector multiplication are introduced for further enhancement in computation. An open-source MATLAB/Simulink-based computational tool, namely the Hybrid2D, is developed to promote the use of the improved CPMs in RTHS as numerical substructures. Numerical simulations are first conducted to verify the accuracy and the computational efficiency of the improved CPMs in Hybrid2D. Subsequently, two RTHS examples are experimentally conducted to demonstrate the effectiveness of the Hybrid2D in enabling RTHS with a higher-fidelity numerical substructure for more complex structural systems.

Optimization of phosphorus wet acid extraction from sewage sludge ashes: Detailed process insight via multi-variate statistical techniques

To tackle the issue of phosphorus (P) supply, Sewage Sludge Ash (SSA) is a promising secondary source. The goal of this research is the optimization of the wet P extraction process in the view of providing valuable insights for the process scale-up. While most of the studies relied on the “one-variable-changed-at-a-time” approach, this work applied a statistical optimization model based on the Design of Experiments technique for investigating the influence of most relevant operating conditions of extraction process on Phosphorus Extraction Efficiency (PEE) with two types of SSA characterized by different levels of alkalinity. Experimental results highlighted the importance of ash characteristics, making some SSA not suitable for cost-effective phosphorus recovery due to the high acid dosage required in extraction process. Type of acid and contact time become significant factors only for Ca-rich ashes, especially when using H2SO4 as extracting agent. Optimal operating conditions (CH2SO4 = 1.3 N, L/S = 20, t = 2 h and CHCl = 2.0 N, L/S = 10, t = 2 h for Ash#1 and CH2SO4/HCl = 0.8 N, L/S = 10 for Ash#2) resulting in high PEE (80–94%) were identified for extraction performed with H2SO4 and HCl as a function of ash characteristics.